Section of Rheumatology, Department of Medicine, West Virginia University School of Medicine, Morgantown.

Abstract

Older men (n = 12) and women (n = 18) 65-80 years of age completed 12 weeks of exercise and took either a placebo or resveratrol (RSV) (500 mg/d) to test the hypothesis that RSV treatment combined with exercise would increase mitochondrial density, muscle fatigue resistance, and cardiovascular function more than exercise alone. Contrary to our hypothesis, aerobic and resistance exercise coupled with RSV treatment did not reduce cardiovascular risk further than exercise alone. However, exercise added to RSV treatment improved the indices of mitochondrial density, and muscle fatigue resistance more than placebo and exercise treatments. In addition, subjects that were treated with RSV had an increase in knee extensor muscle peak torque (8%), average peak torque (14%), and power (14%) after training, whereas exercise did not increase these parameters in the placebo-treated older subjects. Furthermore, exercise combined with RSV significantly improved mean fiber area and total myonuclei by 45.3% and 20%, respectively, in muscle fibers from the vastus lateralis of older subjects. Together, these data indicate a novel anabolic role of RSV in exercise-induced adaptations of older persons and this suggests that RSV combined with exercise might provide a better approach for reversing sarcopenia than exercise alone.

(A) Maximal oxygen uptake (VO2 max) was determined before (Pre) and after (Post) 12 wk of exercise in subjects who consumed a placebo (PL) or resveratrol (RSV) throughout the training period. The data are expressed as absolute oxygen consumed. Maximal oxygen consumption increased in RSV-treated subjects but not in PL-treated subjects after 12 wk of exercise. (B) Representative electron micrographs of vastus lateral muscle sections prepared for electron microscopy before training (i: scale bar = 500 µm) and after 12 wk of exercise training in PL-treated (ii: scale bar = 500 µm) and RSV-treated (iii: scale bar = 500 µm) samples. A lower magnification of post-exercise RSV-treated muscle (iv: scale bar = 800 µm). Examples of apparently morphologically normal mitochondria are shown with the red arrows, and apparently, abnormal mitochondria are shown with blue arrows. (C) mRNA was extracted from biopsies of the vastus lateralis muscles before and after exercise. Subjects took either a PL or RSV for 12 wk and exercised throughout the study. Gene expression was analyzed by PCR array and the global data set were evaluated from 84 genes. Quantitative analysis of genes that were differentially increased in muscles of RSV- vs PL-treated older subjects. The Heat Map provides a visualization of the fold changes in mitochondrial log2 fold gene expression between the selected groups for every gene with the layout for the Heat Map as follows: Layout010203040506070809101112AATP5AATP5BATP51ATP5FATP51ATP5ATP53ATP5ATP5IATP5ATP5JATP5BATP5OCOX4I1COX5ACOX5BCOX6A1COX6A2COX6B1COX6CCOX7A2COX7A2COX7BCOX8ACCYC1NDUFA1NDUFA1NDUFA11NDUFA3NDUFA3NDUFA4NDUFA5NDUFA6NDUFA8NDUFAB1NDUFB1DNDUFB2NDUFB3NDUFB4NDUFB5NDUFB6NDUFB7NDUFB8NDUFB9NDUFC1NDUFC2NDUFS1NDUFS2ENDUFS3NDUFS4NDUFS5NDUFS6NDUFS7NDUFS8NDUFV1NDUFV2NDUFV3PPA1SDHASDHBFSDHCSDHDUQCR11UQCRC1UQCRC2UQCRFS1UQCRHUQCRQARRDC3ASB1CYB561DDNAJB1GEDN1GADD45BHSPA1AHSPA1BLRP5LMitoH1MitoH2_MitoH2_MitoH2_4MitoH2_RNU11SSLC25A25The legend for the color intensity is shown below each heat map.(D) Isokinetic measures of knee extensor muscle torque (Newton-meters), average peak torque, and average power (watts) were obtained on a Biodex at 60°/s. The data represent mean data from the subjects in each group. *p < .05, pre-training vs post-training. **p < .05, RSV vs PL treatment. Pre-PL, before training in the placebo group; Post-PL, after 12 wk of training in the placebo group. Pre-RSV, before training in the resveratrol group; Post-RSV, resveratrol-treated muscles after 12 wk of exercise. *p < .05, pre-training vs post-training.

An example of tissue sections cut from human biopsies. (A) DAPI (blue)-stained nuclei. (B) Pax7-positive satellite cell nuclei are green. (C) The basal lamina of each fiber was identified by a laminin antibody (fuchsia). (D) The overlay of panels A–C. White arrows point to Pax7 nuclei that are located inside the basal lamina. The yellow arrow shows a Pax7 nucleus that is outside of the basal lamina. (E) Total nuclei and satellite cell (Pax7+) content were quantified and expressed per fiber. Nuclei that were outside of the basal lamina were assumed to be of non-myogenic origins. Although Pax7 nuclei that were outside of the basal lamina would be expected to be capable of myogenic contributions, they were not counted as it was not possible to tell if they were migrating from another muscle fiber, or if they would contribute to new de novo muscle fibers. *p < .05, pre-training vs post-training. **p < .05, RSV vs PL treatment. (F) 8-Hydroxyguanosine (8-OHdG)-positive nuclei were identified and quantified if they resided inside the basal lamina as an indication of oxidative damage to skeletal muscle nuclear DNA. *p < .05, pre-training vs post-training. (G) A representative cross-section from a 78-year-old female after 12 wk of exercise training using a PL. (i) DAPI-stained nuclei (blue). (ii) 8-OHdG-positive nuclei are green. (iii) The muscle sarcolemma (red) was indicated by an anti-dystrophin antibody. (iv) The basal lamina was identified by an antibody to laminin and this is shown in white. (v) The overlay of the preceding images is shown. White arrows indicate the same 8-OHdG-positive nuclei in each section. Nuclei that were located inside the basal lamina and were 8-OHdG positive were quantified. Most nuclei that sat between the basal lamina and sarcolemma would be expected to be satellite cells, but these cells were only rarely 8-OHdG positive, whereas more frequently, non-muscle nuclei that resided outside of the basal lamina and sarcolemma were identified as 8-OHdG positive (yellow arrow) but these nuclei were not quantified.